The present invention relates to methods and products for reducing chloride secretion using aromatic organic compounds. In particular the invention relates to methods of treating diarrhea and scours by administering triaryl methane compounds.
Acute and chronic diarrheas represent a major medical problem in many areas of the world. Diarrhea is both a significant factor in malnutrition and the leading cause of death (5,000,000 deaths/year) in children less than five years old. Secretory diarrheas are also a dangerous condition in patients of acquired immunodeficiency syndrome (AIDS) and chronic inflammatory bowel disease (IBD). 16 million travelers to developing countries from industrialized nations every year develop diarrhea, with the severity and number of cases of diarrhea varying depending on the country and area of travel. The major medical consequences of diarrheal diseases include dehydration, acidosis, death and impaired growth.
Diarrhea in barn animals and pets such as cows, pigs and horses, sheep, goats, cats and dogs, also known as scours, is a major cause of death in these animals. Diarrhea can result from any major transition, such as weaning or physical movement. One form of diarrhea is characterized by diarrhea in response to a bacterial or viral infection and generally occurs within the first few hours of the animal""s life.
Although the major consequences of diarrheal diseases are very similar, there are numerous causes of diarrhea. Secretory and exudative diarrhea are primarily caused by bacterial or viral infections. The most common diarrheal causing bacteria is enterotoxogenic E-coli (ETEC) having the K99 pilus antigen. Common viral causes of diarrhea include rotavirus and coronavirus. Other infectious agents include cryptosporidium, giardia lamblia, and salmonella, among others.
The treatment for diarrhea depends on the patient and the infection source. Diarrhea which is found in travelers to industrialized nations (travelers diarrhea) frequently is caused by bacterial pathogens which are acquired through ingestion of fecally contaminated food and/or water. Approximately 50-75% of these cases are attributed to ETEC. Although traveler""s diarrhea is painful, it is generally not life-threatening and often the symptoms last only 3-5 days. The symptoms include urgent diarrhea, abdominal cramps, nausea and fever. The most effective course of treatment for traveler""s diarrhea is the administration of antibiotics in conjunction with oral rehydration. It has been shown that prophylactic administration of antibiotics drastically reduces the number of travelers experiencing symptoms of diarrhea. However, routine administration of antibiotics is not suggested as it may cause resistant strains of a bacteria to develop. Other treatment methods include administration of bismuth subsalicylate, often taken in the form of Pepto-Bismal, diphenoxylate and loperamide.
Diarrhea in AIDS patients is a very serious condition which causes wasting and may be an important factor in the decline of these patients. AIDS patients often develop diarrhea due to enteric infections which their immune system is not capable of fighting off, but AIDS patients may also develop diarrhea by AIDS enteropathy. AIDS enteropathy is a disorder characterized by diarrhea without the involvement of secondary infections. It is caused by the human immunodeficiency virus (HIV) infection of the small bowel mucosal cells and colonic mucosal cells. The most common infective agent causing diarrhea due to enteric infection in AIDS patients in cryptosporidium. The methods for treating diarrhea in AIDS patients include administration of antibiotics and administration of immunoglobulins or an immunoglobulin enriched fraction of bovine colostrum. Colostrum, which is the first milk produced by mammals after birthing is enriched with antibodies.
Acute diarrhea or scours, is a main cause of death in many newborn barn animals such as calves and pigs. Scours is often caused by ETEC with a K99 pilus antigen. Infection with the ETEC causes hypersecretion of fluid and electrolytes. Hypersecretion in turn causes dehydration and pH imbalance which may result in death of the newborn calf or pig.
Newborn barn animals are also susceptible to viral infectious agents causing scours. Infections with rotavirus and coronavirus are common in newborn calves and pigs. Rotavirus infection often occurs within 12 hours of birth. Symptoms of rotaviral infection include excretion of watery feces, dehydration and weakness. Coronavirus which causes a more severe illness in the newborn animals, has a higher mortality rate than rotaviral infection. Often, however, a young animal may be infected with more than one virus or with a combination of viral and bacterial microorganisms at one time. This dramatically increases the severity of the disease.
Generally the best protection for a newborn barn animal from viral or bacterial infection is the consumption of colostrum. If the mother animal has been exposed to these infectious agents then the colostrum will contain antibodies, which are often sufficient to protect the newborn from contracting the diseases. Sometimes, however, this is not sufficient and the animals need further protection. A common method of treatment includes administration of a concentrated colostrum solution or an immunoglobulin fraction isolated from a colostrum solution. This oral treatment may be combined with rehydration salts. Although these methods have improved the morbidity and mortality rate of newborn animals having scours, there still exists a need for more effective treatments.
Certain imidazoles such as clotrimazole are agents which have been used both topically and systemically as antifungals. More recently, studies have identified other uses for such imidazoles. U.S. Pat. No. 5,273,992 revealed that these imidazoles regulate Ca++ activated K+ channels in erythrocytes, and are thus useful in treating sickle cell anemia, which involves the inhibition of potassium transport. These imidazoles have also been found to be effective in inhibiting endothelial and/or vascular smooth muscle cell proliferation. The results of this finding are described in U.S. Pat. No. 5,358,959 and U.S. Ser. No. 08/018,840, which discloses using clotrimazole for treating atherosclerotic and angiogenic conditions, respectively. Nonimidazole metabolites and analogs of the foregoing compounds also have been described as useful in treating the foregoing conditions (see U.S. Ser. Nos. 08/307,874 and 08/307,887).
The present invention provides methods and products for treating diarrhea and scours. It has been discovered that aromatic compounds are effective in treating patients with diarrhea and animals with scours. These compounds are potent inhibitors of secretagogue-stimulated transepithelial electrogenic chloride secretion in intestinal cells.
According to one aspect of the invention, a method for treating diarrhea of diverse etiology is provided. The method involves administering to a subject who is in need of such treatment, an aromatic compound of the invention in an amount effective to inhibit the diarrhea. Preferably the compound is administered orally in conjunction with oral rehydration fluids. The aromatic compounds useful in the invention have the following formula: 
or a pharmaceutically acceptable salt or hydrate thereof, wherein: n is 0, 1, 2, 3 or 4; p is 0 or 1; X is absent, (C1-C3)alkyl, (C1-C3)alkenyl, (C1-C3)alkynyl, SCH2, OCH2, or NOCH2, Y is C, N, P, Si or Ge; Rxe2x80x2 is absent, -halo, xe2x80x94R, xe2x80x94OR, xe2x80x94SR, xe2x80x94NR2, xe2x80x94ONR2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94C(O)R, xe2x80x94C(S)R, xe2x80x94C(O)OR, xe2x80x94C(S)OR, xe2x80x94C(O)SR, xe2x80x94C(S)SR, xe2x80x94C(O)NR2, xe2x80x94C(S)NR2, xe2x80x94C(O)NR(OR), xe2x80x94C(S)NR(OR), xe2x80x94C(O)NR(SR), C(S)NR(SR), xe2x80x94CH(CN)2, xe2x80x94CH[C(O)R]2, xe2x80x94CH[C(S)R]2, xe2x80x94CH[C(O)OR]2, xe2x80x94CH[C(S)OR]2, xe2x80x94CH[C(O)SR]2, xe2x80x94CH[C(S)SR]2, aryl, or heteroaryl; Ar1 is aryl, substituted aryl, heteroaryl, (C5-C8)cycloalkyl or (C5-C8)heterocycloalkyl; Ar2 is aryl or substituted aryl; Ar3 is aryl, substituted aryl, biaryl, biphenyl, bibenzyl, or heteroaryl other than imidazole, nitroimidazole and triazole; each R is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C1-C6)alkenyl, substituted (C1-C6)alkenyl(C1-C6)alkynyl, substituted (C1-C6)alkynyl, and (C1-C6)alkoxy; the aryl substituents are each independently selected from the group consisting of -halo, trihalomethyl, xe2x80x94R, xe2x80x94R1, xe2x80x94OR1, xe2x80x94SR1, NR12, xe2x80x94NO2, xe2x80x94CN, xe2x80x94C(O)R1, xe2x80x94C(S)R1, xe2x80x94C(O)OR1, xe2x80x94C(S)OR1, xe2x80x94C(O)SR1 and xe2x80x94C(S)SR1; the alkyl, alkenyl and alkynyl substituents are each independently selected from the group consisting of -halo, xe2x80x94R1, xe2x80x94OR1, xe2x80x94SR1, N(R1)2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94C(O)R1, xe2x80x94C(S)R1, xe2x80x94C(O)OR1, xe2x80x94C(S)OR1, xe2x80x94C(O)SR1, xe2x80x94C(S)SR1, aryl, xcex3-butyrolactonyl, pyrrolidinyl and succinic anhydridyl; and each R1 is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl and (C1-C6)alkynyl.
In a preferred embodiment p=0, X is absent, (C1-C3)alkyl, (C1-C3)alkenyl, (C1-C3) or alkynyl; Rxe2x80x2 is absent, xe2x80x94halo, xe2x80x94R, xe2x80x94OR, xe2x80x94SR, xe2x80x94NR2, xe2x80x94ONR2, xe2x80x94NO2, xe2x80x94CN, xe2x80x94C(O)R, xe2x80x94C(S)R, xe2x80x94C(O)OR, xe2x80x94C(S)OR, xe2x80x94C(O)SR, xe2x80x94C(S)SR, xe2x80x94C(O)NR2, xe2x80x94C(S)NR2, xe2x80x94C(O)NR(OR), xe2x80x94C(S)NR(OR), xe2x80x94C(O)NR(SR), C(S)NR(SR), xe2x80x94CH(CN)2, xe2x80x94CH[C(O)R]2, xe2x80x94CH[C(S)R]2, xe2x80x94CH[C(O)OR]2, xe2x80x94CH[C(S)OR]2, xe2x80x94CH[C(O)SR]2, xe2x80x94CH[C(S)SR]2, or aryl; Ar1 is aryl, substituted aryl, heteroaryl other than imidazole, nitroimidazole and triazole, heteroarylium other than imidazolium, nitroimidazolium and triazolium, (C5-C8) cycloalkyl or (C5-C8)heterocycloalkyl; Ar2 is aryl or substituted aryl; and Ar3 is aryl, substituted aryl, biaryl, or heteroaryl other than imidazole, nitroimidazole and triazole.
In a preferred embodiment the aromatic compounds are those of formula (I), except that the compounds are not any compound encompassed by formula (II): 
wherein n=0-3; wherein p=0 or 1; wherein X is selected from the group consisting of (CH2)m(m=0,1,2, or 3), CHxe2x95x90CH, Cxe2x89xa1C, SCH2, OCH2, and NOCH2; wherein Rxe2x80x2 is selected from the group consisting of H, OH, SH, NO2, CN, CHO, ONH2, CCH, CORxe2x80x3, CO2H, CO2Rxe2x80x3, ORxe2x80x3, SRxe2x80x3, NRxe2x80x3Rxe2x80x3, CONRxe2x80x3Rxe2x80x3, heteroaryl, and CONRxe2x80x3(OCH3); wherein Ar1 is selected from the group consisting of phenyl, substituted phenyl, and heteroaryl; wherein Ar2 is selected from the group consisting of phenyl and substituted phenyl; wherein Ar3 is selected from the group consisting of phenyl, substituted phenyl, biphenyl, bibenzyl, and naphthyl; wherein the phenyl substituent is selected from the group consisting of Cl, F, Br, I, R, ORxe2x80x3, SRxe2x80x3, NO2, CN, CF3, NRxe2x80x3Rxe2x80x3, and CO2R; wherein R is selected from the group consisting of straight chain alkyl of Cz(z=1-5), substituted straight chain alkyl of Cz(z=1-5) branched alkyl of Cz(z=1-5), and substituted branched alkyl of Cz(z=1-5); wherein the alkyl substituent is selected from the group consisting of Cl, Br, F, I, OH, OCH3, SH, SCH3, NH2, NHCH3, and N(CH3)2; and wherein Rxe2x80x3 is selected from the group consisting of hydrogen and R.
In one embodiment of the invention the foregoing aromatic compounds may be administered in combination with other non-formula (I) anti-diarrheal agents. In another embodiment the aromatic compounds may be administered in combination with other non-formula (I) anti-scours agents.
According to one embodiment of the invention the subject in need of such treatment is a subject who has symptoms of diarrhea or scours. In another embodiment of the invention, the subject in need of such treatment is a subject at risk of developing diarrhea or scours.
In general diarrhea is a secretory disorder, which is caused by at least one of several mechanisms. In one embodiment the diarrhea is an exudative form of diarrhea; In one embodiment the diarrhea is a nonexudative form of diarrhea; In another embodiment the diarrhea is a decreased absorption form of diarrhea; In another embodiment the diarrhea is a non-decreased absorption form of diarrhea; In yet another embodiment the diarrhea is a secretory form of diarrhea. In yet another embodiment the diarrhea is a nonsecretory form of diarrhea. In still another embodiment the diarrhea is a noninflammatory form of diarrhea.
According to another aspect of the invention, pharmaceutical preparations are provided. These pharmaceutical preparations include the aromatic compounds of the invention together with an anti-diarrheal agent. In one embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above. In another embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above but do not include the compounds of formula (II). In yet another embodiment, the aromatic compounds useful according to the invention have the above-disclosed general formula (I), but wherein Rxe2x80x2 and Ar1 do not include imidazoles. Preferably the pharmaceutical composition of the invention may be administered orally.
According to another aspect of the invention, veterinary preparations are provided. These veterinary preparations include the aromatic compounds useful according to the invention together with an anti-scours preparation. In one embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above. In another embodiment, the aromatic compounds useful according to the invention have the general formula (I) provided above but do not include the compounds of formula (II). In yet another embodiment, the aromatic compounds useful according to the invention have the above-disclosed general formula (I), but wherein Rxe2x80x2 and Ar1 do not include imidazoles.
The invention also provides the aromatic compounds of the invention in the manufacture of a medicament for the treatment of diarrhea. In one embodiment, the aromatic compounds of the invention have the above-disclosed general formula, but do not include clotrimazole. In another embodiment, the aromatic compounds useful in the manufacture of a medicament for the treatment of diarrhea have the above-disclosed general formula, but wherein Rxe2x80x2 and Ar1 do not include imidazoles.
The invention also provides the aromatic compounds of the invention in the manufacture of a medicament for the treatment of scours. In one embodiment, the aromatic compounds of the invention have the general formula (I). In another embodiment, the aromatic compounds useful in the manufacture of a medicament for the treatment of scours have the general formula (I) provided above but do not include the compounds of formula (II).
In embodiments of each of the methods or products of the invention the aromatic compounds have the following the structural formula (III): 
wherein:
n is 0, 1, 2, 3 or 4;
R1 is xe2x80x94H, xe2x80x94OR, xe2x80x94SR, xe2x80x94CN, xe2x80x94C(O)R, xe2x80x94C(O)OR, xe2x80x94C(O)NR2, xe2x80x94CH[C(O)R]2 or xe2x80x94CH[C(O)OR]2;
R2 is xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94OR, xe2x80x94SR, xe2x80x94C(O)R or xe2x80x94C(O)NR2;
R2xe2x80x2 is xe2x80x94H or xe2x80x94NO2;
R3 is xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, xe2x80x94OR or xe2x80x94SR;
R4 is xe2x80x94H or xe2x80x94NR2;
R4xe2x80x2 is xe2x80x94H, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I; and
each R is independently selected from the group consisting of xe2x80x94H, (C1-C6) alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl or (C1-C6)alkoxy.
In another preferred embodiment, the aromatic compounds of the invention are compounds having the structural formula (IV): 
wherein:
X is absent or xe2x80x94Cxe2x89xa1Cxe2x80x94;
Y is C, P, Si or Ge;
n is 0, 1, 2, 3 or 4;
Ar1 is phenyl, substituted phenyl, cycloalkyl or heteroarylium other than imidazolium, nitroimidazolium or triazolium;
Ar3 is phenyl, naphthyl, piperidyl or cyclohexyl;
R1 is xe2x80x94R, xe2x80x94OR, xe2x80x94SR, xe2x80x94CN, xe2x80x94NR2, xe2x80x94ONR2, xe2x80x94C(O)R, xe2x80x94C(O)OR, xe2x80x94C(O)NR2, xe2x80x94CH[C(O)R]2, xe2x80x94CH[C(O)OR]2, (C1-C6)alkyl, (C1-C6)alkenyl, (C1xe2x80x94C6)alkynyl, cyclopenta-2,4-diene-1-ylidene or phenyl;
each of R2, R3 and R4 is independently selected from the group consisting of xe2x80x94H, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94OR, xe2x80x94SR, xe2x80x94NR2, xe2x80x94NO2, xe2x80x94C(O)R, xe2x80x94C(O)OR, xe2x80x94C(O)NR2, trihalomethyl, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl and phenyl;
each R is independently selected from the group consisting of xe2x80x94H, halo, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C1-C6)alkenyl, substituted (C1-C6)alkenyl, (C1-C6)alkynyl, substituted (C1-C6)alkynyl and (C1-C6)alkoxy;
the alkyl, alkenyl or alkynyl substituents are each independently selected from the group consisting of aryl, xe2x80x94C(O)OR, pyrrolidinyl, butyrolactonyl, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I and xe2x80x94CN; and
the phenyl substituents are each independently xe2x80x94R.
In another preferred embodiment, the aromatic compounds of the invention are compounds having the formula (V): 
or pharmaceutically acceptable salts or hydrates thereof, wherein:
n is 0, 1, 2, 3 or 4;
R is xe2x80x94H, xe2x80x94OR, xe2x80x94SR, xe2x80x94CN, xe2x80x94C(O)R, xe2x80x94C(S)R, xe2x80x94C(O)OR, xe2x80x94C(S)OR, xe2x80x94C(O)SR, xe2x80x94C(S)SR, xe2x80x94C(O)NR2, xe2x80x94C(S)NR2, xe2x80x94CH[C(O)R]2, xe2x80x94CH[C(S)R]2, xe2x80x94CH[C(O)OR]2, xe2x80x94CH[C(S)OR]2, xe2x80x94CH[C(O)SR]2, xe2x80x94CH[C(S)SR]2;
R2 is xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I;
R3 is xe2x80x94R, xe2x80x94OR or xe2x80x94SR;
R4 is xe2x80x94H or xe2x80x94NR2;
R4xe2x80x2 is xe2x80x94H, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I; and
each R is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl and (C1-C6)alkoxy.
In another preferred embodiment, the compounds of the invention are those of formula (V), with the provisos that (i) when n is 0 and R1 is xe2x80x94H or xe2x80x94OH, R3 is other than xe2x80x94H; and (ii) when n is 0 and R1 is xe2x80x94H, R3 is other than xe2x80x94OH.
In another preferred embodiment, the compounds of the invention are those of formula (V), with the proviso that when n is 0 and R1 is xe2x80x94C(O)NH2, R2 is other than xe2x80x94F.
In another preferred embodiment, the aromatic compounds of the invention are compounds having the formula (VI): 
or pharmaceutically acceptable salts or hydrates thereof, wherein:
n is 0, 1, 2, 3 or 4;
R1 is xe2x80x94NR2, xe2x80x94C(O)R, xe2x80x94C(S)R, xe2x80x94C(O)NRxe2x80x22 or xe2x80x94C(S)NRxe2x80x22;
R2 is xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I;
R3 is xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I;
R4 is xe2x80x94F, xe2x80x94Cl, xe2x80x94Br or xe2x80x94I;
each R is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl and (C1-C6)alkoxy; and
each Rxe2x80x2 is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl and (C1-C6)alkoxy.
In another preferred embodiment, the compounds of the invention are compounds having the formula (VII): 
or pharmaceutically acceptable salts or hydrates thereof, wherein:
n is 0, 1, 2, 3 or 4;
Ar1 is phenyl or cyclohexyl;
R, is xe2x80x94NR2, xe2x80x94CH[C(O)OR]2, xe2x80x94CH[C(S)OR]2, xe2x80x94CH[C(O)SR]2, xe2x80x94CH[C(S)SR]2, xe2x80x94C(O)NR2 or xe2x80x94C(S)NR2; and
each R is independently selected from the group consisting of xe2x80x94H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl and (C1-C6)alkoxy.
Each of the limitations of the invention can encompass various embodiments of the invention. It is, therefore, anticipated that each of the limitations of the invention involving any one element or combinations of elements can be included in each aspect of the invention.